Screening, Isolation, Structure Elucidation Of Natural Sources Nuroactive Components

Alzheimer's diease is a kind of neurodegenerative disease mainly affecting both cognition and behavior with unknown etiology. PC 12 cell is a immature neuron cell line sources of rat adrenal pheochromocytoma cells, has been widely used in pathological research and drugs screening of nervous system diease. Traditional Chinese medicine has been used to treat various human ailments for thousands years in China. Traditional Chinese medicine contain a lot of bioactive components, such as polysaccharide,terpenes,flavonoids and so on, have been reported to exhibit various pharmaceutical effects including regulating immune, antitumor, antibiotic, neurtrophin etc. At present, some taditional Chinese medicine have been reported to exhibit neurotophin. But previous studies focused mainly on promoting PC 12 cell differentiation. Studying on tracking on neuroactive components of taditional Chinese medicine's isolation has rarely been reported.In this study, promoting PC12 cell neurite outgrowth model were used to screen 98 kinds of Taditional Chinese medicine,and isolation,purification,determination the active compounds. Reserch shows that:1. Extracts of 98 taditional Chinese medicines were screened by PC 12 cell model, isolation of Tremella fuciformis lipophilic extracts have a certain function to promote PC 12 cell neurite outgrowth, the effect is obvious, and isolation of Dysosma lipophilic extracts was tracked on in complete hibition NGF active component in PC 12 cell.2. Studying on tracking on neuroactive components of T. fuciformis:through the first silica gel open column, got 7 samples, the best neurotive site ZY-II-20-6, that neurite outgrowth rate is 33% at the concentration of 1μg/mL:isolated by the second silica gel open column, got 5 samples, the best neurotive site ZY-Ⅱ-25-4, that neurite outgrowth rate is 30% at the concentration of 1μg/mL; and then once ODS open column. ZY-Ⅱ-35-3 is the best activity better than other 4 samples, that neurite outgrowth rate is 42% at the concentration of 1μ/mL; after the semi prepareative HPLC, got a active compound A-6 at 3μmol/L concentration, the neurite outgrowth rate is 56%. Measuring A-6 to 1H-NMR,13C-NMR,DEPT1351(?),ESIMS,H-Ucosy,HMBC,HSQC, found A-6 is 3-O-β-D- Glucopyranosyl-22E,24R-5α,8α-epidioxyergosta-6,22-diene. This compound is the first found in T. fuciformis.3. Dysosma lipophilic extracts at 3μmol·L-1have a obvious inhibition effect to PC12 cell neurite outgrowth, through the silica gel open column, got 7 parts,B-3 and B-5 quiescing at room temperature, it separated crystal, named B-8 and B-9. B-9 inhibited the PC12 cells neurite outgrowth, the ratio is 100% at 0.3μmol·L-1; treatd PC12 cells with 40 ng/mL NGF and 3μmol·L-1 B-9. found the ratio is 100% too. Measuring B-9 to 1H-NMR,13C-NMR,ESIMS,found it is podophyllotoxin. It rarely been reported to inhibit NGF activity.